Bottom Line:
Rg1 significantly increased the activities of antioxidant enzymes (SOD, GSH-Px and CAT) and reduced MDA levels in liver tissues.Furthermore, Rg1 significantly increased the expression and nuclear translocation of Nrf2 that regulated the expression of many antioxidant enzymes.Knockdown of Nrf2 gene diminished these actions of Rg1 in CCl4-treated HSCs in vitro.

fig7: Nrf2 playing a key role in the mechanism of anti-hepatic fibrosis by Rg1. (A) Effect of Rg1 on prolieration in HSCs. Primary HSCs were incubated with Rg1 and CCl4 (10 mmol/L) for 24 h. Data represent mean±SD of 6 separate experiments. (B) Nrf2 siRNA inhibits Rg1-induced translocation of Nrf2. Cells were treated with 1 μmol/L Rg1 for 24 h after control or Nrf2 siRNA transfection for 48 h. Protein expression of Nrf2 was detected by Western blotting. Representative blots were from three independent experiments. The content of MDA (C), GPX (D), PCIII (E), and HA (F) in the cell cultured supernatant and the α-SMA expression in HSCs (G). Cells were treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h after control or Nrf2 siRNA transfection for 48 h; non-siRNA groups were cultured for 48 h and then treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h. The data were expressed as mean±SD. n=6. Statistical evaluation was performed using t-test. bP<0.05, cP<0.01 vs control. eP<0.05, fP<0.01 vs CCl4 alone. hP<0.05, iP<0.01 vs Rg1+CCl4 group.

Mentions:
The CCl4-induced HSC proliferation was inhibited by Rg1 (10−7, 10−6 mol/L) compared with that of the CCl4 only group (P<0.05, P<0.01, respectively; Figure 7A). As 10−6 mol/L of Rg1 showed a better and more stable effect, this concentration was chosen for the following study.

fig7: Nrf2 playing a key role in the mechanism of anti-hepatic fibrosis by Rg1. (A) Effect of Rg1 on prolieration in HSCs. Primary HSCs were incubated with Rg1 and CCl4 (10 mmol/L) for 24 h. Data represent mean±SD of 6 separate experiments. (B) Nrf2 siRNA inhibits Rg1-induced translocation of Nrf2. Cells were treated with 1 μmol/L Rg1 for 24 h after control or Nrf2 siRNA transfection for 48 h. Protein expression of Nrf2 was detected by Western blotting. Representative blots were from three independent experiments. The content of MDA (C), GPX (D), PCIII (E), and HA (F) in the cell cultured supernatant and the α-SMA expression in HSCs (G). Cells were treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h after control or Nrf2 siRNA transfection for 48 h; non-siRNA groups were cultured for 48 h and then treated with 1 μmol/L Rg1 and CCl4 (10 mmol/L) or DMSO for 24 h. The data were expressed as mean±SD. n=6. Statistical evaluation was performed using t-test. bP<0.05, cP<0.01 vs control. eP<0.05, fP<0.01 vs CCl4 alone. hP<0.05, iP<0.01 vs Rg1+CCl4 group.

Mentions:
The CCl4-induced HSC proliferation was inhibited by Rg1 (10−7, 10−6 mol/L) compared with that of the CCl4 only group (P<0.05, P<0.01, respectively; Figure 7A). As 10−6 mol/L of Rg1 showed a better and more stable effect, this concentration was chosen for the following study.

Bottom Line:
Rg1 significantly increased the activities of antioxidant enzymes (SOD, GSH-Px and CAT) and reduced MDA levels in liver tissues.Furthermore, Rg1 significantly increased the expression and nuclear translocation of Nrf2 that regulated the expression of many antioxidant enzymes.Knockdown of Nrf2 gene diminished these actions of Rg1 in CCl4-treated HSCs in vitro.